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首页> 外文期刊>Journal of bacteriology >Global View of the Clostridium thermocellum Cellulosome Revealed by Quantitative Proteomic Analysis
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Global View of the Clostridium thermocellum Cellulosome Revealed by Quantitative Proteomic Analysis

机译：定量蛋白质组学分析揭示了热纤梭菌纤维素体的整体视图。

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A metabolic isotope-labeling strategy was used in conjunction with nano-liquid chromatography-electrospray ionization mass spectrometry peptide sequencing to assess quantitative alterations in the expression patterns of subunits within cellulosomes of the cellulolytic bacterium Clostridium thermocellum, grown on either cellulose or cellobiose. In total, 41 cellulosomal proteins were detected, including 36 type I dockerin-containing proteins, which count among them all but three of the known docking components and 16 new subunits. All differential expression data were normalized to the scaffoldin CipA such that protein per cellulosome was compared for growth between the two substrates. Proteins that exhibited higher expression in cellulosomes from cellulose-grown cells than in cellobiose-grown cells were the cell surface anchor protein OlpB, exoglucanases CelS and CelK, and the glycoside hydrolase family 9 (GH9) endoglucanase CelJ. Conversely, lower expression in cellulosomes from cells grown on cellulose than on cellobiose was observed for the GH8 endoglucanase CelA; GH5 endoglucanases CelB, CelE, CelG; and hemicellulases XynA, XynC, XynZ, and XghA. GH9 cellulases were the most abundant group of enzymes per CipA when cells were grown on cellulose, while hemicellulases were the most abundant group on cellobiose. The results support the existing theory that expression of scaffoldin-related proteins is coordinately regulated by a catabolite repression type of mechanism, as well as the prior observation that xylanase expression is subject to a growth rate-independent type of regulation. However, concerning transcriptional control of cellulases, which had also been previously shown to be subject to catabolite repression, a novel distinction was observed with respect to endoglucanases.
机译：代谢同位素标记策略与纳米液相色谱-电喷雾电离质谱肽测序结合使用，以评估生长于纤维素分解细菌上的纤维素分解细菌的纤维素体中亚基表达模式的定量变化。纤维素或纤维二糖。总共检测到41种纤维素蛋白，其中包括36种含I型dockerin的蛋白，除三个已知的对接组分和16个新的亚基外，其他全部计数。将所有差异表达数据相对于支架蛋白CipA标准化，从而比较每个纤维素小体的蛋白质在两种底物之间的生长。与纤维二糖生长的细胞相比，在纤维素生长的细胞中的纤维素酶体中表达更高的蛋白是细胞表面锚蛋白OlpB，外切葡聚糖酶CelS和CelK，以及糖苷水解酶家族9（GH9）内切葡聚糖酶CelJ。相反，对于GH8内切葡聚糖酶CelA，在纤维素上生长的细胞在纤维素小体中的表达低于纤维二糖。 GH5内切葡聚糖酶CelB，CelE，CelG;半纤维素酶XynA，XynC，XynZ和XghA。当细胞在纤维素上生长时，GH9纤维素酶是每个CipA中最丰富的酶，而半纤维素酶是纤维二糖上最丰富的酶。结果支持现有的理论，即支架蛋白相关蛋白的表达受分解代谢物抑制类型的机制协调调节，以及先前观察到的木聚糖酶表达受生长速率非依赖性调节类型的影响。然而，关于纤维素酶的转录控制，先前也已经显示其受到分解代谢物抑制，关于内切葡聚糖酶观察到新的区别。

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